Quick Protocol for Monarch® PCR & DNA Cleanup Kit ( 5 μg ) ( NEB # T1030 )
Dilute sample with DNA Cleanup Binding Buffer according to the table below .
Mix well by pipetting up and down or flicking the tube .
Do not vortex .
A sample volume of 20–100 μl is recommended .
For smaller samples , TE can be used to adjust the volume .
For diluted samples larger than 800 μl , load a portion of the sample , proceed with step 2 , and then repeat as necessary .
Sample TypeRatio of Binding Buffer :
SampleExampledsDNA > 2 kb ( plasmids , gDNA ) 2 : 1200 μl : 100 μldsDNA < 2 kb ( some amplicons , fragments ) 5 : 1500 μl : 100 μlssDNA ( cDNA , M13 ) 7 : 1700 μl : 100 μl .
Insert column into collection tube and load sample onto column .
Spin for 1 minute at 16,000 x g , then discard flow - through .
Re - insert column into collection tube .
Add 200 μl DNA Wash Buffer ( with ethanol added ) and spin for 1 minute at 16,000 x g .
Discarding flow - through is optional .
Repeat Step 3 .
( Step 3 : Re - insert column into collection tube .
Add 200 μl DNA Wash Buffer and spin for 1 minute at 16,000 x g .
Discarding flow - through is optional ) .
Transfer column to a clean 1.5 ml microfuge tube .
Use care to ensure that the tip of the column does not come into contact with the flow - through .
If in doubt , re - spin for 1 minute .
Add ≥ 6 μl of DNA Elution Buffer to the center of the matrix .
Wait for 1 minute , then spin for 1 minute at 16,000 x g to elute the DNA .
